Conjunctive Use of Various Adjuvant and Fusion Protein Which Composed of M2e and N P Genes of Avian Influenza Virus,and the Influence on Immunogenicity
- VernacularTitle:禽流感病毒M2、NP融合蛋白与多种佐剂的联合运用及对免疫原性影响
- Author:
Hai XU
;
Hong-Yan HOU
;
Bi-Hua DENG
;
Qi-Sheng ZHENG
;
Ji-Bo HOU
;
- Publication Type:Journal Article
- Keywords:
Avian Influenza Virus Antigen epitope Adjuvent Immunogenicity
- From:
China Biotechnology
2006;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Based on the gene sequence of AIV matrix protein 2(M2) and two cytotoxic T-lymphocyte epitopes derived from nucleoprotein,the prokaryotic expression vector pET-3M2e-NP1-NP2 was constructed.The target gene was expressed in the solvable form in BL21(DE3) when induced with 1.0 mmol/L IPTG and Western blot analysis showed that the expression product had good immunogencity.Purified fusion protein was mixed with various amount of adjuvant,including Freund's,Vash oil and chitosan,and then immunized 20-day-old chicken by intramuscular injection and boosted 3weeks later.Blood samples were collected weekly following the primary vaccination.The anti-M2e antibody was detected with ELISA method with the synthesized peptide as coated antigen;the neutralizing ability of anti-serum was evaluated on MDCK cell line and chick embryo,the CD4+ and CD8+ T lymphocyte amounts in peripheral blood of immunized chicken was measured by flow cytometry.Results showed that the fusion protein can induce immunological reaction,and the antibody can bind with the viral M2 protein that expressed on the surface of MDCK cells.Flow cytometry result showed that CD4+ and CD8+ T lymphocyte in peripheral blood increased obviously following immunization(P
